1.Differences in deltamethrin resistance and kdr gene mutation in Culex tritaeniorhynchus population in and outside the Yellow Sea wetland
Xiao-er ZHANG ; Zhi-ming WU ; Ye TIAN ; Qian CUI ; Yu-qian JI ; Huan WANG ; Shu-juan YANG ; Yi-chao ZHAO ; Yu WANG ; Hua-yu YIN ; Yu DING ; Guo-jin YAN ; Min-sen ZHAO ; Shou-gang ZHANG ; Bing-dong SONG ; Hong-na CHEN ; Jian GAO ; Wei-fang YANG ; Yu-fu ZHANG ; Hui LIU ; Hong-liang CHU
Acta Parasitologica et Medica Entomologica Sinica 2026;33(2):101-107
Objective To gain insights into the biological characteristics of different populations of Culex tritaeniorhynchus within and around the Yellow Sea wetland from the perspective of the occurrence of resistance, we investigated the levels of resistance to deltamethrin and kdr gene mutation in the wetland and its peripheral areas. Methods Specimens were collected from Cx. tritaeniorhynchus populations at two monitoring sites in the Rare Bird National Nature Reserve and Tiaozi Ni Wetland Scenic Area, and also from two populations in Yancheng City and the Liuhe District of Nanjing, and the resistance of these mosquitoes to deltamethrin was determined using the CDC biotest bottle method. For each concentration of deltamethrin assessed, a random subset of exposed specimens was selected for amplification of the kdr gene fragment, followed by Sanger sequencing to identify and analyze resistance-associated mutations. Results The LC50 levels of deltamethrin among mosquitoes from the four populations in Luhe, Yancheng, the Rare Bird National Nature Reserve and the Tiaozi Ni Wetland Scenic Area were 2.048 5, 7.798 2, 3.473 3, and 17.695 5 mg/mL, respectively, with corresponding concentrations of deltamethrin ranging from 0.005 to 5.000,0.050 to 50.000,0.050 to 25.000 and 0.050 to 50.000 mg/mL, respectively. Furthermore, the ranges of the KT50 values were 11.76-107.43, 67.05-216.30,29.77-107.43 and 28.40-329.51 min; the 1-h knockdown rates were 34.58%-99.15%, 9.52%-43.80%, 55.09%-73.01%, and 10.09%-68.07%; and the 24-h mortality rates were 12.15%-67.52%,9.52%-79.56%,13.17%-82.21%, and 11.01%-78.99%, respectively. With respect to kdr gene mutation, we assayed a total of 63,70,59, and 57 mosquitoes for the four populations, for which we detected L1014F mutation frequencies of 14.29%, 35.00%, 20.34%, and 31.58%, respectively, with a majority of these mutations being heterozygous for resistance. In addition, five adult mosquitoes were identified has having synonymous mutations at site 1011[i. e. , AAT(asparagine)mutation to AAC(asparagine)]. Conclusions Our findings revealed the clear resistance of Cx. tritaeniorhynchus to deltamethrin in the Yancheng region of the Yellow Sea wetland, and the resistance phenotype and kdr frequency of Cx. tritaeniorhynchus in the wetland environment were comparable to those of Cx. tritaeniorhynchus in the wetland environment, thereby indicating that the resistance of different populations of Cx. tritaeniorhynchus was homogeneous under the pressure of different insecticide selection within and around the wetland. However, the underlying mechanisms need to be further studied.
2.Effect of different phosphorus application on morphological traits, active ingredients and rhizosphere soil microbial community of Polygala tenuifolia.
Huan GUO ; Tong WEI ; Wen-Hua CUI ; Huan SHI ; Fu-Ying MAO ; Xian GU ; Yun-Sheng ZHAO ; Xiao-Feng LIANG
China Journal of Chinese Materia Medica 2025;50(14):3898-3908
To investigate the effects of phosphorus fertilizer on the morphological traits, active ingredients and rhizosphere soil microbial community of Polygala tenuifolia. The phosphorus fertilizer was calculated in terms of P_2O_5. Five treatments were set up: 0(CK), 17(P1), 34(P2), 51(P3), and 68(P4) kg per Mu(1 Mu≈667 m~2). A randomized block design was adopted. Samples of P. tenuifolia and its rhizosphere soil were collected under different superphosphate fertilizer treatments. Illumina high-throughput sequencing was used to analyze the rhizosphere soil microbial community, 9 morphological traits were measured and the content of 11 active ingredients were determined. The results showed that the whole plant weight, shoot fresh weight, root weight, and root peel thickness were the highest under P1 treatment, increasing by 34.41%, 38.80%, 39.21%, and 3.17% respectively compared to CK. Under P2 treatment, the plant height, stem diameter, root thickness, and core thickness were significantly higher than CK. Phosphorus fertilizer had a significant impact on the content of tenuifolin, sibiricose A5, sibiricose A6, arillanin A, 3,6'-disinapoyl sucrose, and polygalaxanthone Ⅲ. Correlation analysis results showed that the relative abundance of Arthrobacter, Bacillus, norank_f_Vicinamibacteraceae, norank_o_Vicinamibacterales, MND1 and other bacteria, as well as the relative abundance of Neocosmospora, Paraphoma and other fungi were positively correlated with root diameter, wood core diameter, the whole plant weight, root weight, shoot fresh weight of P. tenuifolia. Bacillus, Neocosmospora, Subulicystidium were significantly positively correlated with oligosaccharides such as 3,6'-disinapoyl sucrose, sibiricose A5、sibiricose A6、glomeratose A、arillanin A and tenuifoliside C. Arthrobacter, Humicola, Aspergillus, Paraphoma were positively correlated with tenuifolin and norank_f_Vicinamibacteraceae, norank_o_Vicinamibacterales, Fusarium were positively correlated with polygalaxanthone Ⅲ. Evidently, appropriate phosphorus application is conducive to the growth and quality improvement of P. tenuifolia, and can increase the abundance of beneficial microorganisms in the soil.
Rhizosphere
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Phosphorus/pharmacology*
;
Soil Microbiology
;
Polygala/anatomy & histology*
;
Fertilizers/analysis*
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Bacteria/metabolism*
;
Soil/chemistry*
;
Microbiota/drug effects*
;
Plant Roots/metabolism*
3.The application of robot-assisted positioning in total hip arthroplasty for patients with coronal pelvic tilt
Yinggang ZHENG ; Huan XIAO ; Libo HAO ; Jun FU ; Yongjian LIANG ; Zhiyuan LI ; Te LIU ; Chi XU
Chinese Journal of Orthopaedics 2025;45(17):1104-1110
Objective:To explore the advantages of robotic-arm assisted total hip arthroplasty (rTHA) in acetabular component positioning and lower limb length assessment in patients with severe pelvic coronal tilt.Methods:A retrospective analysis was conducted on the data of 122 patients with unilateral end-stage hip disease and coronal pelvic tilt angle >3° who underwent total hip arthroplasty (THA) at the First Medical Center of PLA General Hospital from June 2022 to December 2023. Among them, 44 patients underwent rTHA, and 78 underwent manual THA (mTHA). The rTHA group included 18 males and 26 females, with an average age of 60.5±9.3 years; the mTHA group included 41 males and 37 females, with an average age of 58.5±8.4 years. Compare the differences in the anteversion angle, abduction angle, pelvic tilt angle, leg length discrepancy (LLD) of the acetabular prosthesis, and the proportions of patients with LLD>0.5 cm and >1 cm between the two groups of patients after surgery. Calculate the proportion of outlier rates of acetabular abduction angle (<30° or >45°), and proportions within Callanan's safe zone. The early efficacy of the hip joint was evaluated by using the modified Harris score and joint range of motion.Results:All patients were followed up for 6 to 12 months, with an average of 8 months. All the surgical incisions of the patients achieved primary healing. Postoperative comparisons showed no statistically significant differences in acetabular abduction angle (39.5°±3.3° vs. 38.4°±7.3°) or anteversion angle (20.7°±1.6° vs. 19.7°±1.6°) between rTHA and mTHA groups ( P>0.05). However, pelvic tilt angle [2.5° (1.1°, 3.6°) vs. 3.5° (2.3°, 5.9°)] showed a statistically significant difference ( U=4.371, P=0.008). The rTHA group exhibited smaller absolute LLD [0.2 (0.1, 0.4) cm vs. 0.5 (0.2, 0.5) cm] and lower proportions of LLD >0.5 cm [14% (6/44) vs. 49% (38/78)] and >1 cm [2% (1/44) vs. 18% (14/78)], with statistical significance ( P<0.05). The rTHA group had a lower outlier rate for acetabular abduction angle (<30°or >45°) compared to the mTHA group [2% (1/44) vs. 33% (26/78)], with statistical significance (χ 2=10.388, P<0.001). Taking the Callanan safety zone as the standard, the proportion of acetabular cups within the safe zone was significantly higher in the rTHA group (98%, 43/44) compared to the mTHA group (67%, 52/78), with a statistically significant difference (χ 2=13.998, P<0.001). The modified Harris score and hip joint range of motion in the mTHA group increased from 47.6±6.6 points and 83° (73°, 88°) before the operation to 83.5±11.2 points and 118° (110°, 122°) at the last follow-up, respectively. The rTHA group increased from 46.5±9.2 points and 79° (71°, 90°) before the operation to 85.0±12.5 points and 124° (116°, 130°) at the last follow-up. The differences in the modified Harris score and hip joint range of motion between the two groups before the operation and at the last follow-up were statistically significant ( P<0.05). However, there was no statistically significant difference between the groups at the last follow-up ( P>0.05). No THA-related complications occurred during follow-up period. Conclusion:For patients with end-stage hip joint diseases with coronal tilt exceeding 3°, robotic-assisted technology significantly improves the accuracy of acetabular component placement during THA and offers better control of postoperative LLD.
4.Genetic diversity analysis of oxacillinase in 241 clinical isolates of Pseudomonas aeruginosa
Yuelong LI ; Jingyi ZHANG ; Yubing FU ; Meiqing SUN ; Beibei MIAO ; Xinyi GONG ; Xiao HAN ; Huan XING ; Pengfang GAO ; Jiachen LI ; Yating TANG ; Xinya FAN ; Yanlei GE ; Haijian ZHOU ; Juan LI ; Aiying DONG
Chinese Journal of Preventive Medicine 2025;59(7):1004-1012
Objective:To analyze the carriage status, subtype distribution and flanking gene sequence characteristics of oxacillinases (OXA enzyme) in 241 clinical strains of Pseudomonas aeruginosa, and assess their roles in the drug resistance of Pseudomonas aeruginosa and ability to horizontally transfer across species. Methods:Clinical P. aeruginosa isolates were collected from four hospitals in Sanya, Tangshan, Zhangjiakou, and Beijing. The prevalence of oxacillinases and their flanking gene sequences was analyzed by whole-genome sequencing (NGS) and bioinformatic approaches. Results:A total of 241 isolates of P. aeruginosa were gathered, and 35 blaOXA subtypes were identified through screening of 252 blaOXA genes. These genes were classified into three subfamilies: blaOXA-50-like (241, 95.6%), blaOXA-1-like (9, 3.6%) and blaOXA-10-like (2, 0.8%). Among these, 11 subtypes (11, 31.4%) were novel blaOXA subtypes. Nine of these belonged to the blaOXA-50-like subfamily and were designated as blaOXA-1244, blaOXA-1245, blaOXA-1246, blaOXA-1250, blaOXA-1252, blaOXA-1253, blaOXA-1254, blaOXA-1255, and blaOXA-1256. The remaining two belonged to the blaOXA-10-like subfamily and were named blaOXA-1247 and blaOXA-1248. Compared to the amino acid sequence of OXA-10, the newly identified subtype OXA-1247 exhibited a mutation at position 117, where a valine was replaced by a leucine. This change was thought to improve the enzyme′s ability to hydrolyze carbapenems. In the analysis of the flanking sequences of the blaOXA genes, Class I integrons were identified in four bacterial strains. The variable regions of these integrons carried three distinct patterns of resistance gene cassettes: aac( 6′) -Ib-blaOXA-1247-ant( 3′′) -Ia, aac( 6′) -Ib-blaOXA-1248 and aac( 6′) -Ib- blaIMP-45-blaOXA-1-catB3. Among these, the strain BJ2326 carried a class I integron that was connected to the downstream IS CR1 element to form a composite class I integron structure, additionally carrying the resistance gene blaPER-1. Out of the 223 non-wild-type P. aeruginosa strains, 127 strains exhibited non-wild-type profiles to the four beta-lactam antibiotics MEM, CAZ, FEP, and TZP, with the combination of MEM+CAZ+FEP being the most prevalent, representing 57.0% of the total. Conclusions:The blaOXA genes in 241 clinical P. aeruginosa strains showed diversity. Some blaOXA genes had a co-transfer risk with the metallo-β-lactamase resistance gene blaIMP-45. Among the 11 newly discovered blaOXA subtypes, the new subtype OXA-1247 may have carbapenemase activity and potential for horizontal transfer.
5.FOXQ1 inhibits DNA damage-induced apoptosis in colorectal cancer cells by promoting p53 deacetylation
Gui-song YANG ; Huan-jie CHEN ; Gui-liang MA ; Fu-gang WANG ; Xiao-lei MA ; Hong QI
Chinese Journal of Current Advances in General Surgery 2025;28(4):266-270
Objective:To investigate the effect of FOXQ1 expression on chemoresistance in colorectal cancer and analyze its regulatory role in SIRT1 expression and p53 deacetylation under DNA damage response(DDR)condi-tions.Methods:qRT-PCR was used to detect FOXQ1 mRNA expression levels in SW620 cells and SW620 cells stimulated with cisplatin(CDDP).Lentiviral vectors were constructed for FOXQ1 overexpression and RNA interference.The cells were divided into three groups:FOXQ1 overexpression group(oe-FOXQ1),FOXQ1 RNA interference group(sh-FOXQ1),and a control group transfected with an empty vector(NC).The half-maximal inhibitory concentration(IC50)of CDDP in each group was determined using the CCK-8 assay.Apoptosis level and cell viability were assessed using the Annexin V-APC/7-ADD apoptosis detection kit and Calcein/PI staining.Western blot analysis was performed to evaluate the effect of FOXQ1 on SIRT1 expression and acetylated p53 levels.The SIRT1 pathway inhibitor(S)-Selisi-stat was introduced to observe changes in p53 acetylation levels.Results:Compared to normal colon tissues,FOXQ1 expression was significantly upregulated in SW620 cells(P<0.05),and low-dose CDDP stimulation further en-hanced its expression(P<0.05).After 24 hours of CDDP treatment,the IC50 values for the oe-FOXQ1,sh-FOXQ1,and NC groups were 58.3 μmol/L,36.4 μmol/L,and 43.7 μmol/L,respectively,with statistically significant differences among the groups(P<0.05).Compared to the NC group,the oe-FOXQ1 group showed a decrease in late apoptotic cell count(P<0.05),while the sh-FOXQ1 group exhibited an increase(P<0.05).Cytotoxic fluorescence staining re-vealed that the proportion of cell death was lower in the oe-FOXQ1 group and higher in the sh-FOXQ1 group com-pared to the NC group(P<0.05).Protein expression analysis showed that FOXQ1 and SIRT1 levels were higher in the oe-FOXQ1 group and lower in the sh-FOXQ1 group compared to the NC group(P<0.05).FOXQ1 overexpression pro-moted p53 deacetylation,while the addition of the SIRT1 pathway inhibitor(S)-Selisistat restored p53 acetylation levels(P<0.05).Conclusion:FOXQ1 promotes p53 deacetylation by upregulating SIRT1 expression,thereby inhibiting DDR-induced apoptosis.
6.FOXQ1 inhibits DNA damage-induced apoptosis in colorectal cancer cells by promoting p53 deacetylation
Gui-song YANG ; Huan-jie CHEN ; Gui-liang MA ; Fu-gang WANG ; Xiao-lei MA ; Hong QI
Chinese Journal of Current Advances in General Surgery 2025;28(4):266-270
Objective:To investigate the effect of FOXQ1 expression on chemoresistance in colorectal cancer and analyze its regulatory role in SIRT1 expression and p53 deacetylation under DNA damage response(DDR)condi-tions.Methods:qRT-PCR was used to detect FOXQ1 mRNA expression levels in SW620 cells and SW620 cells stimulated with cisplatin(CDDP).Lentiviral vectors were constructed for FOXQ1 overexpression and RNA interference.The cells were divided into three groups:FOXQ1 overexpression group(oe-FOXQ1),FOXQ1 RNA interference group(sh-FOXQ1),and a control group transfected with an empty vector(NC).The half-maximal inhibitory concentration(IC50)of CDDP in each group was determined using the CCK-8 assay.Apoptosis level and cell viability were assessed using the Annexin V-APC/7-ADD apoptosis detection kit and Calcein/PI staining.Western blot analysis was performed to evaluate the effect of FOXQ1 on SIRT1 expression and acetylated p53 levels.The SIRT1 pathway inhibitor(S)-Selisi-stat was introduced to observe changes in p53 acetylation levels.Results:Compared to normal colon tissues,FOXQ1 expression was significantly upregulated in SW620 cells(P<0.05),and low-dose CDDP stimulation further en-hanced its expression(P<0.05).After 24 hours of CDDP treatment,the IC50 values for the oe-FOXQ1,sh-FOXQ1,and NC groups were 58.3 μmol/L,36.4 μmol/L,and 43.7 μmol/L,respectively,with statistically significant differences among the groups(P<0.05).Compared to the NC group,the oe-FOXQ1 group showed a decrease in late apoptotic cell count(P<0.05),while the sh-FOXQ1 group exhibited an increase(P<0.05).Cytotoxic fluorescence staining re-vealed that the proportion of cell death was lower in the oe-FOXQ1 group and higher in the sh-FOXQ1 group com-pared to the NC group(P<0.05).Protein expression analysis showed that FOXQ1 and SIRT1 levels were higher in the oe-FOXQ1 group and lower in the sh-FOXQ1 group compared to the NC group(P<0.05).FOXQ1 overexpression pro-moted p53 deacetylation,while the addition of the SIRT1 pathway inhibitor(S)-Selisistat restored p53 acetylation levels(P<0.05).Conclusion:FOXQ1 promotes p53 deacetylation by upregulating SIRT1 expression,thereby inhibiting DDR-induced apoptosis.
7.The application of robot-assisted positioning in total hip arthroplasty for patients with coronal pelvic tilt
Yinggang ZHENG ; Huan XIAO ; Libo HAO ; Jun FU ; Yongjian LIANG ; Zhiyuan LI ; Te LIU ; Chi XU
Chinese Journal of Orthopaedics 2025;45(17):1104-1110
Objective:To explore the advantages of robotic-arm assisted total hip arthroplasty (rTHA) in acetabular component positioning and lower limb length assessment in patients with severe pelvic coronal tilt.Methods:A retrospective analysis was conducted on the data of 122 patients with unilateral end-stage hip disease and coronal pelvic tilt angle >3° who underwent total hip arthroplasty (THA) at the First Medical Center of PLA General Hospital from June 2022 to December 2023. Among them, 44 patients underwent rTHA, and 78 underwent manual THA (mTHA). The rTHA group included 18 males and 26 females, with an average age of 60.5±9.3 years; the mTHA group included 41 males and 37 females, with an average age of 58.5±8.4 years. Compare the differences in the anteversion angle, abduction angle, pelvic tilt angle, leg length discrepancy (LLD) of the acetabular prosthesis, and the proportions of patients with LLD>0.5 cm and >1 cm between the two groups of patients after surgery. Calculate the proportion of outlier rates of acetabular abduction angle (<30° or >45°), and proportions within Callanan's safe zone. The early efficacy of the hip joint was evaluated by using the modified Harris score and joint range of motion.Results:All patients were followed up for 6 to 12 months, with an average of 8 months. All the surgical incisions of the patients achieved primary healing. Postoperative comparisons showed no statistically significant differences in acetabular abduction angle (39.5°±3.3° vs. 38.4°±7.3°) or anteversion angle (20.7°±1.6° vs. 19.7°±1.6°) between rTHA and mTHA groups ( P>0.05). However, pelvic tilt angle [2.5° (1.1°, 3.6°) vs. 3.5° (2.3°, 5.9°)] showed a statistically significant difference ( U=4.371, P=0.008). The rTHA group exhibited smaller absolute LLD [0.2 (0.1, 0.4) cm vs. 0.5 (0.2, 0.5) cm] and lower proportions of LLD >0.5 cm [14% (6/44) vs. 49% (38/78)] and >1 cm [2% (1/44) vs. 18% (14/78)], with statistical significance ( P<0.05). The rTHA group had a lower outlier rate for acetabular abduction angle (<30°or >45°) compared to the mTHA group [2% (1/44) vs. 33% (26/78)], with statistical significance (χ 2=10.388, P<0.001). Taking the Callanan safety zone as the standard, the proportion of acetabular cups within the safe zone was significantly higher in the rTHA group (98%, 43/44) compared to the mTHA group (67%, 52/78), with a statistically significant difference (χ 2=13.998, P<0.001). The modified Harris score and hip joint range of motion in the mTHA group increased from 47.6±6.6 points and 83° (73°, 88°) before the operation to 83.5±11.2 points and 118° (110°, 122°) at the last follow-up, respectively. The rTHA group increased from 46.5±9.2 points and 79° (71°, 90°) before the operation to 85.0±12.5 points and 124° (116°, 130°) at the last follow-up. The differences in the modified Harris score and hip joint range of motion between the two groups before the operation and at the last follow-up were statistically significant ( P<0.05). However, there was no statistically significant difference between the groups at the last follow-up ( P>0.05). No THA-related complications occurred during follow-up period. Conclusion:For patients with end-stage hip joint diseases with coronal tilt exceeding 3°, robotic-assisted technology significantly improves the accuracy of acetabular component placement during THA and offers better control of postoperative LLD.
8.Genetic diversity analysis of oxacillinase in 241 clinical isolates of Pseudomonas aeruginosa
Yuelong LI ; Jingyi ZHANG ; Yubing FU ; Meiqing SUN ; Beibei MIAO ; Xinyi GONG ; Xiao HAN ; Huan XING ; Pengfang GAO ; Jiachen LI ; Yating TANG ; Xinya FAN ; Yanlei GE ; Haijian ZHOU ; Juan LI ; Aiying DONG
Chinese Journal of Preventive Medicine 2025;59(7):1004-1012
Objective:To analyze the carriage status, subtype distribution and flanking gene sequence characteristics of oxacillinases (OXA enzyme) in 241 clinical strains of Pseudomonas aeruginosa, and assess their roles in the drug resistance of Pseudomonas aeruginosa and ability to horizontally transfer across species. Methods:Clinical P. aeruginosa isolates were collected from four hospitals in Sanya, Tangshan, Zhangjiakou, and Beijing. The prevalence of oxacillinases and their flanking gene sequences was analyzed by whole-genome sequencing (NGS) and bioinformatic approaches. Results:A total of 241 isolates of P. aeruginosa were gathered, and 35 blaOXA subtypes were identified through screening of 252 blaOXA genes. These genes were classified into three subfamilies: blaOXA-50-like (241, 95.6%), blaOXA-1-like (9, 3.6%) and blaOXA-10-like (2, 0.8%). Among these, 11 subtypes (11, 31.4%) were novel blaOXA subtypes. Nine of these belonged to the blaOXA-50-like subfamily and were designated as blaOXA-1244, blaOXA-1245, blaOXA-1246, blaOXA-1250, blaOXA-1252, blaOXA-1253, blaOXA-1254, blaOXA-1255, and blaOXA-1256. The remaining two belonged to the blaOXA-10-like subfamily and were named blaOXA-1247 and blaOXA-1248. Compared to the amino acid sequence of OXA-10, the newly identified subtype OXA-1247 exhibited a mutation at position 117, where a valine was replaced by a leucine. This change was thought to improve the enzyme′s ability to hydrolyze carbapenems. In the analysis of the flanking sequences of the blaOXA genes, Class I integrons were identified in four bacterial strains. The variable regions of these integrons carried three distinct patterns of resistance gene cassettes: aac( 6′) -Ib-blaOXA-1247-ant( 3′′) -Ia, aac( 6′) -Ib-blaOXA-1248 and aac( 6′) -Ib- blaIMP-45-blaOXA-1-catB3. Among these, the strain BJ2326 carried a class I integron that was connected to the downstream IS CR1 element to form a composite class I integron structure, additionally carrying the resistance gene blaPER-1. Out of the 223 non-wild-type P. aeruginosa strains, 127 strains exhibited non-wild-type profiles to the four beta-lactam antibiotics MEM, CAZ, FEP, and TZP, with the combination of MEM+CAZ+FEP being the most prevalent, representing 57.0% of the total. Conclusions:The blaOXA genes in 241 clinical P. aeruginosa strains showed diversity. Some blaOXA genes had a co-transfer risk with the metallo-β-lactamase resistance gene blaIMP-45. Among the 11 newly discovered blaOXA subtypes, the new subtype OXA-1247 may have carbapenemase activity and potential for horizontal transfer.
9.Characterization of genomic islands and virulence factors of clinical isolates of Burkholderia pseudomallei in Hainan Province,China
Xiao-Ying FU ; Huan LI ; Sha LI ; Li-Cheng WANG ; Chong-Zhen WANG ; Yuan-Li LI ; Hai CHEN ; Xiong ZHU
Chinese Journal of Zoonoses 2024;40(4):359-368,390
The genomic island(GI)characteristics and virulence factor differences of clinical isolates of Burkholderia pseudomallei in Hainan Province,China were analyzed to provide a scientific basis for diagnosis and treatment of melioidosis.In total,52 B.pseudomallei isolates were collected for detection of virulence-related GIs by PCR.The whole genome sequence annotation format file was submitted on Islandviwer 4 platform,and the genomes of the same species and close relatives were added for comparison.Two algorithms,SIGI-HMM and IslandPath-DIMOB,were integrated to predict GIs and sequence a-lignments were conducted to identify specific GIs and differences in virulence factors.The genomes of 52 clinical strains could be divided into three branches based on evolutionary distance,with 82.69%(43/52)of strains concentrated in branch 1.In to-tal,828 GIs were identified among the 52 B.pseudomallei genomes,which formed 157 GI clusters based on sequence similari-ty.GIs accounted for 2.05%-6.38%of the genome content.While GI clusters 1 and 2 were present in all strains,a total of 84(53.50%)GI clusters only clustered within a single genome isolate.Of 10 GI likely specific clusters,five were from the same genus,two from another genus,and three with uncertain origins.Moreover,25 GI clusters were associated with virulence,which included eight shared by B.pseudomallei BP76 and BP169,which had the highest number of virulence-associated GIs among all isolates.O the 52 B.pseudomallei isolates,variations were identified in the virulence genes fhaB1,fhaB2,BPSL1661,cheY1,wzM,tssH-5/clpV,tssA-5,boaA,and boaB.Comparisons of these findings with clinical isolates from Thailand and Australia showed that B.pseudomallei isolates from Hainan had significant differences in the sequences of boaA,boaB,cheY1,and chbp.Additionally,fhaB1,fhaB3,and bimA displayed significant variations specifically within the Australian isolates.B.pseudomallei GI was conserved and specific to Hainan.The identification of specific GI and virulence factors was useful to clarify the source of horizontal gene transfer and differences in virulence at the molecular level.
10.Dynamic changes of neuronal cells at different time points following cerebral ischemia-reperfusion injury in rats
Xu-Huan ZOU ; Rui LAN ; Xue-Qin FU ; Wei-Wei WANG ; Man-Man WANG ; Chen TANG ; Shuang LIU ; Hong-Yu LI ; Xiao-Ming SHEN
Chinese Pharmacological Bulletin 2024;40(6):1056-1066
Aim To investigate the dynamic changes of neuronal cells at different time points following acute cerebral ischemia-reperfusion injury by establishing a model of brain ischemia-reperfusion injury.Methods Thirty male Sprague-Dawley(SD)rats were ran-domly divided into six groups:sham group and cere-bral ischemia-reperfusion injury(IR)groups at differ-ent time points.Focal cerebral ischemia-reperfusion injury model was established using the middle cerebral artery occlusion(MCAO)technique.The Longa sco-ring method was used to assess neurobehavioral scores in rats.After successful model preparation,routine paraffin sections were made,and TUNEL staining and immunohistochemistry staining with NeuN antibody were performed to observe cell apoptosis and neuronal cell survival,respectively.Immunohistochemistry stai-ning was also performed to investigate the changes in glial fibrillary acidic protein(GFAP)as a marker for astrocytes,ionized calcium-binding adapter molecule 1(IBA-1)as a marker for microglia,and CD31 as a marker for endothelial cells at different time points.Results No significant changes were observed in neu-ronal cells of the sham group at different time points.In the cerebral ischemia-reperfusion injury groups,cell apoptosis was activated at IR3h and increased in quan-tity with morphological damage as time progressed.Ne-uN+neurons showed signs of ischemic injury after IR3h,with abnormal cell morphology.From 12 h,Ne-uN+neurons decreased in a time-dependent manner and reached their peak severity at 24 h.GFAP+astro-cytes decreased significantly after IR3h,while poorly labeled GFAP+astrocytes increased at IR 6 h and al-most disappeared in the infarcted area at 24 h and 48 h.The number of IBA-1+microglia-positive cells de-creased at IR3h,and their volume increased at IR6h.Microglial cell death was observed in the infarcted area at IR12h.CD31+endothelial cells around the infarc-ted cortex and striatum increased significantly after IR3h and persisted until 48 h.Conclusions After cerebral ischemia-reperfusion injury,the number of ap-optotic cells increases with the prolongation of time,and NeuN+neurons exhibit the most severe damage at 24 h.GFAP+astrocytes and microglial cells gradually die over time.The number of CD31+endothelial cells increases significantly around the infarcted cortex and striatum after 3 h of reperfusion and persists until 48 h.


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